LOCUS       sequence-147695-        3319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Supplemental Figure 3. Integration Site 7..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3319)
  AUTHORS   Perez-Pinera P, Han N, Cleto S, Cao J, Purcell O, Shah KA, Lee K, 
            Ram R, Lu TK
  TITLE     Synthetic biology and microbioreactor platforms for programmable 
            production of biologics at the point-of-care.
  JOURNAL   Nat Commun. 2016 Jul 29;7:12211. doi: 10.1038/ncomms12211.
  PUBMED    27470089
REFERENCE   2  (bases 1 to 3319)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2016-07-29"; volume: "7:12211. doi"; pages: " 
            10.1038/ncomms12211"
COMMENT     Sequence Label: PP69
FEATURES             Location/Qualifiers
     source          1..3319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      158..405
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(174..192)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(480..1068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(560..579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1159..1192)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    1802..1849
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     promoter        2165..2578
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2585..2632
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(2657..3319,1..153)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(2657..3319,1..153)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2730..2749)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
       61 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
      121 cagtttcatt tgatgctcga tgagtttttc taacacatca tgtaattagt tatgtcacgc
      181 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
      241 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
      301 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
      361 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac
      421 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      481 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      541 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      601 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      661 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      721 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      781 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      841 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      901 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      961 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1021 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1081 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1141 atgcatgaga tcagatctga agttcctata ctttctagag aataggaact tcaagcttca
     1201 acgatgccaa aagggtataa taagcgtcat ttgcagcatt gtgaagaaaa ctatgtggca
     1261 agccaagcct gcgaagaatg tattttaagt ttgactttga tgtattcact tgattaagcc
     1321 ataattctcg agtatctatg attggaagta tgggaatggt gatacccgca ttcttcagtg
     1381 tcttgaggtc tcctatcaga ttatgcccaa ctaaagcaac cggaggagga gatttcatgg
     1441 taaatttctc tgacttttgg tcatcagtag gctcgaactg tgagactatc tcggttatga
     1501 cagcagaaat gtccttcttg gagacagtaa atgaagtccc accaataaag aaatccttgt
     1561 catcaggaac aaacttcttg tttcgaactt tttcggtgcc ttgaactata atcatgagat
     1621 tattggaaac caccagaatc gaatataaaa ggcgaacacc tttcccaatt ttggtttctc
     1681 ctgacccaaa gactttaaat ttaatttatt tgtccctatt tcaatcaatt gaacaactat
     1741 ttcggctgga cggcgacgta aacggccaca agtttatggc cgtgatgacc tgtgtcttcg
     1801 tggtttgtct ggtcaaccac cgcggtctca gtggtgtacg gtacaaaccc aaagcagcac
     1861 gacacggcaa ctacaagacc cgcgccgagg gcatgttccc caaagcgata ccacttgaag
     1921 cagtggtact gcttgtgggt acactctgcg ggtgtgaagt tcgagggcga caccctggtg
     1981 aaccgcatcg agctgaaggg catcttccaa ctcgcttaat tgcgagtttt tatttcgttt
     2041 atttcaatta aggtaactaa aaaactcctt ttacacatga agcagcacga cttcttcaag
     2101 tccgccatgc ccgaaaaacg cctcttcaga gtacagaaga ttaagtgaga ccttcgtttg
     2161 tgcggatccc ccacacacca tagcttcaaa atgtttctac tcctttttta ctcttccaga
     2221 ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa
     2281 tttcccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga
     2341 aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac
     2401 gtttcttttt cttgaaaatt tttttttttg atttttttct ctttcgatga cctcccattg
     2461 atatttaagt taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct
     2521 attacaactt tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaaggg
     2581 cggtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     2641 gaactaaacc atggtaatga gccatattca acgggaaacg tcttgctcta ggccgcgatt
     2701 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2761 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2821 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2881 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2941 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
     3001 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3061 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3121 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3181 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3241 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3301 tgatgttgga cgagtcgga
//
